Anellated dihydropyridines and the use thereof for the production of pharmaceutical preparation

ABSTRACT

Compound of general formula I ##STR1## wherein A is a benzo or thieno group; 
     R 1  is (C 4-6 )cycloalkyl, (C 4-6 )cycloalkyl-(C 1-5 )alkyl or ##STR2## R 2 , m, R 3 , R 4 , R and u are defined as in the specification, and pharmaceutical preparations containing this compound and the new pharmaceutical uses thereof.

This is a continuation of application Ser. No. 08/249,822 filed May 26,1994 now abandoned.

The invention relates to anellated dihydropyridinoacetic acidderivatives, processes for preparing them and pharmaceuticalcompositions containing these compounds.

Dihydroisoquinolines are known from EP-A 37 934. The compounds mentionedtherein have a cardiotonic activity and have an activity component whichincreases contractility and influences blood pressure. They have beenproposed for improving circulation of the blood through the tissues andfor improving the supply of oxygen to the tissues. These possible usesare based on the vascular activity of the compounds. EP-A 251 194describes how carbocyclically and heterocyclically anellateddihydropyridines have a cardioprotective activity and constitute anentirely new type of Ca-antagonistic compounds.

The present invention relates to new carbocyclically andheterocyclically anellated dihydropyridines, and the pharmaceutical useof these compounds and new pharmaceutical uses for thedihydroisoquinolines known from EP-A-37 934. These new uses are based ontheir antiproliferative effect and their activity in the treatment ofulcerative colitis and Crohn's disease.

One aspect of the invention consists in the use of a compound of generalformula I ##STR3## wherein A denotes a benzo, indolo or thieno group;wherein, if A is benzo, m is 2 or 3 (preferably 2, the two R² groupsbeing in positions 6 and 7), and the substituents R² independently ofeach other may represent hydroxy, (C₁₋₄)alkoxy, benzyloxy, halogen (F,Cl, Br, I), (C₁₋₄)alkyl, methanesulphonyloxy or methanesulphonamido, ortwo adjacent substituents R² together represent --O--CH₂ --O-- or--O--CH₂ --CH₂ --O--; and if A is indolo or thieno, m is zero;

R₁ denotes (C₄₋₆)cycloalkyl, (C₄₋₆)cycloalkyl(C₁₋₅)alkyl or ##STR4## R³and R⁴ independently of each other denote (a) hydrogen,

(b) branched or unbranched C₃₋₆ -alkenyl,

(c) branched or unbranched C₃₋₆ -alkynyl or

(d) branched or unbranched C₁₋₁₂ -alkyl, whilst the alkyl may besubstituted by

hydroxy,

di(C₁₋₄)alkylamino,

furyl,

pyridyl,

pyrrolidinyl, N-methylpyrrolidinyl,

morpholino,

indolyl,

nitrilo,

thienyl,

adamantyl,

cyclohexyl,

phenoxy,

naphthyloxy or phenyl, whilst this phenyl or the phenyl contained in thephenoxy group may be mono-, di- or trisubstituted by hydroxy,(C₁₋₄)alkoxy, benzyloxy, halogen (F, Cl, Br, I), CF₃, N₃, (C₁₋₄)alkyl,adamantyl, --SO₂ NH₂, --NHCOCH₃, --NHSO₂ CH₃ or CH₃ SO₂ O-- or by thebridge --O--CH₂ --O--;

or R³ denotes hydrogen and R⁴ denotes cyclohexyl, phenyl, fluorophenyl,pyridyl or N-benzylpiperidyl;

or R³ and R⁴ together with the nitrogen atom to which they are boundrepresent

pyrrolidinyl,

piperidinyl,

morpholinyl, thiomorpholinyl, the group ##STR5## or piperazinyl, whilstthe piperazinyl ring may optionally be N-substituted by methyl,unsubstituted phenyl, mono- or di(C₁₋₄)alkoxyphenyl, pyrimidinyl,phenyl(C₁₋₄)alkyl or ##STR6## R denotes C₁₋₄ -alkyl, hydroxy, --N₃,halogen (F, Cl, Br, I), CF₃, C₁₋₄ -alkoxy or --COH and

u denotes 0, 1, 2 or 3;

A further aspect of the invention consists in the following newcompounds: of general formula ##STR7## wherein R¹, R³ and R⁴ are asdefined above; and new compounds of general formula I ##STR8## wherein Adenotes a benzo, indolo or thieno group; wherein, if A is benzo, m is 2or 3 (preferably 2, the two R² groups being in positions 6 and 7), andthe substituents R² independently of each other may represent hydroxy,(C₁₋₄)alkoxy, benzyloxy, halogen (F, Cl, Br, I), (C₁₋₄)alkyl,methanesulphonyloxy or methanesulphonamido, or two adjacent substituentsR² together represent --O--CH₂ --O-- or --O--CH₂ --CH₂ --O--; and if Ais indolo or thieno, m is zero;

if A is indolo or thieno, m is zero;

R₁ denotes (C₄₋₆)cycloalkyl, (C₄₋₆)cycloalkyl(C₁₋₅)alkyl or ##STR9## R³and R⁴ independently of each other denote (a) hydrogen,

(b) branched or unbranched C₃₋₆ -alkenyl,

(c) branched or unbranched C₃₋₆ -alkynyl or

(d) branched or unbranched C₁₋₁₂ -alkyl, whilst the alkyl may besubstituted by

hydroxy,

(C₁₋₄)alkoxy,

di(C₁₋₄)alkylamino,

furyl,

pyridyl,

pyrrolidinyl, N-methylpyrrolidinyl,

morpholino,

indolyl,

nitrilo,

thienyl,

adamantyl,

cyclohexyl,

phenoxy,

naphthyloxy or phenyl, whilst this phenyl or the phenyl contained in thephenoxy group may be mono-, di- or trisubstituted by hydroxy,(C₁₋₄)alkoxy, benzyloxy, halogen (F, Cl, Br, I), CF₃, N₃, (C₁₋₄)alkyl,adamantyl, --SO₂ NH₂, --NHCOCH₃, --NHSO₂ CH₃ or CH₃ SO₂ O-- or by thebridge --O--CH₂ --O--;

or R³ denotes hydrogen and R⁴ denotes cyclohexyl, phenyl, fluorophenyl,pyridyl or or R³ and R⁴ together with the nitrogen atom to which theyare bound represent

pyrrolidinyl,

piperidinyl, morpholinyl, thiomorpholinyl, the group ##STR10## orpiperazinyl, whilst the piperazinyl ring may optionally be N-substitutedby methyl, unsubstituted phenyl, mono- or di(C₁₋₄)alkoxyphenyl,pyrimidinyl, phenyl(C₁₋₄)alkyl or ##STR11## R denotes C₁₋₄ -alkyl,hydroxy, --N₃, halogen (F, Cl, Br, I), CF₃, C₁₋₄ -alkoxy or --COH and

u denotes 0, 1, 2 or 3.

Another aspect of the invention consists in individual new compoundswhich come under the general definition of earlier patent applications.These compounds are contained, together with other compounds, in Tables13 to 20, particularly Tables 14, 16 and 20. The following compounds ofthis group should be particularly mentioned:

New compounds of general formula I of the formula ##STR12## wherein NR³R⁴ has one of the following meanings

    __________________________________________________________________________                                       Structure                                  __________________________________________________________________________     ##STR13##                         I                                           ##STR14##                         I                                           ##STR15##                         I                                           ##STR16##                         I + II                                      ##STR17##                         I                                           ##STR18##                         I                                           ##STR19##                                                                     ##STR20##                                                                    HNCH.sub.2 -1-Adam.                                                            ##STR21##                                                                     ##STR22##                                                                     ##STR23##                                                                     ##STR24##                                                                     ##STR25##                                                                     ##STR26##                         I                                           ##STR27##                         I                                           ##STR28##                         I                                           ##STR29##                         I                                           ##STR30##                         I                                          __________________________________________________________________________    X                                  Structure                                  __________________________________________________________________________     ##STR31##                         I                                           ##STR32##                         II                                          ##STR33##                         I                                          NHCH.sub.2 -Adam(1)                II                                          ##STR34##                         I                                           ##STR35##                                                                     ##STR36##                                                                    __________________________________________________________________________

Compounds of formula I form tautomers of formula II ##STR37##

The tautomers can be separated by known methods, e.g. by columnchromatography or selective reduction (NaBH₄ or catalytic reduction).

The compounds of formula II may be present in cis- and/or trans-form:##STR38##

The invention also comprises the physiologically acceptable salts of theabove new compounds with acids, bases and complexing agents.

The new compounds have valuable therapeutically useful properties. Theycan be used as cardioprotective agents, as cerebroprotective agents(particularly for the treatment of patients who have suffered a strokeor are in danger of suffering a stroke), and as agents for treatingchronic inflammatory processes (e.g. bronchial asthma and arthritis).Furthermore, these compounds can be used as agents with anantiproliferative effect and as agents for treating ulcerative colitisand Crohn's disease.

In the definitions used in the text, the radicals and groups may beidentical or different, i.e. if one of the above-mentioned substituentsoccurs several times in one particular molecule, the meaning can befreely selected on each occasion within the scope of the range ofdefinitions.

The term alkyl refers particularly to C₁₋₆ -alkyl and C₁₋₄ -alkylradicals which may in turn be substituted or, as alkyl radicals, arepart of a functional group such as alkoxy or alkylthio. The alkylradicals include methyl, ethyl, n-propyl, isopropyl, n-butyl, sec.butyl,isobutyl, tert.butyl radicals and the various isomeric pentyl and hexylradicals such as isopentyl, neopentyl, n-pentyl and the n-hexyl radical.

The above definition thus applies even when the alkyl radical is in turnsubstituted and/or is itself part of an alkoxyalkyl, alkoxycarbonyl,alkoxy, alkylthio, alkylsulphonyl, monoalkylamino, alkylmethyl,alkylthiomethyl, dialkylamino group or the alkyl radical is bound as asubstituent to an aromatic heterocyclic or carbocyclic system.

The halogens are fluorine, chlorine, bromine and iodine, preferablyfluorine, chlorine:and bromine and, to a lesser extent, iodine.

C₃₋₆ -cycloalkyl means cyclopropane, cyclobutane, cyclopentane andcyclohexane. C₅₋₆ -cycloalkenes are for example cyclopentene,cyclohexene and cyclohexadiene.

The C₂ - and C₃ -acyl radicals represent the acetyl and propionylradicals.

C₃₋₆ -alkynes are the isomeric hexynes, pentynes, butynes and propynes,preferably propargyl.

The C₃₋₆ -alkenes are the isomeric hexenes, pentenes, butenes andpropenes, preferably allyl.

Examples of unsaturated heterocycles include, inter alia:

furan, pyran, pyrrole, pyrazole, imidazole, pyridine, pyrazine,pyrimidine, pyridazine, thiophene, thiazole, oxazole, 1,2,4-triazole,1,2,3-triazole, 1,2,4-triazine, 1,3,5-triazine, indole.

Examples of 5- or 6-membered, wholly or partially saturated monocyclicheterocycles include inter alia:

imidazolidine, pyrazolidine, pyrrolidine, piperidine, piperazine,morpholine, thiomorpholine, tetrahydrofuran, tetrahydrothiophene,1,4-dioxine, imidazoline, pyrazoline, pyrroline, etc.

A. Of the compounds of formula I wherein R¹ is (C₄₋₆)cycloalkyl or(C₄₋₆)cycloalkyl(C₁₋₅)alkyl, the following may be mentioned as ofparticular interest:

Compounds wherein

R³ and R⁴ independently of each other denote (a) hydrogen, (b) C₁₋₈-alkyl, C₂₋₃ -alkenyl or -alkynyl (wherein the alkyl may be substitutedby hydroxy, (C₁₋₄)alkoxy, di(C₁₋₄)alkylamino, furyl, pyrrolidinyl,morpholinyl, pyridinyl or the group ##STR39## wherein B, R⁵ and q aredefined as hereinafter) (d) dimethylamino, (f) phenyl, (g) morpholinylor (h) pyridyl, whilst R³ and R⁴ cannot simultaneously representhydrogen, dimethylamino or di(C₁₋₄)alkylaminomethyl;

or R³ and R⁴ together with the nitrogen atom to which they are bounddenote a pyrrolidinyl, morpholinyl or piperazinyl group, whilst thepiperazinyl ring may optionally be N-substituted by unsubstitutedphenyl, mono- or di(C₁₋₄)alkoxyphenyl, pyrimidinyl or phenyl(C₁₋₄)alkyl;

more particularly wherein R³ and/or R⁴ denotes unsubstituted phenyl,fluorophenyl, morpholino or 2- or 3-pyridyl;

wherein R³ and/or R⁴ denotes (C₁₋₄)alkyl, preferably methyl or ethyl; or##STR40## wherein R³ and/or R⁴ denotes (C₂ or C₃)alkyl, which issubstituted by hydroxy, methoxy, dimethylamine, furyl, morpholino,pyrrolidinyl or pyridinyl;

wherein R³ is hydrogen.

Moreover, of these compounds (I), special mention should be made ofthose wherein R³ is hydrogen and R⁴ is a substituted alkyl of formulaVII ##STR41## wherein q' is 0, 1 or 2; R⁶ and R⁷ independently of eachother denote hydrogen or (C₁₋₅)alkyl or together with the carbon atom towhich they are bound denote a 5- or 6-membered carbocycle;

B denotes phenyl or thienyl;

R⁵ denotes (C₁₋₄)alkyl, halogen (F, Cl, Br, I), hydroxy, (C₁₋₄)alkoxy,amino, thiomethyl, methanesulphonyloxy or methanesulphonamido, or twoadjacent R⁵ substituents together denote --O--CH₂ --O-- or --O--CH₂--CH₂ --O-- and q' denotes 0, 1, 2 or 3, if B is phenyl, and q' denotes0, 1, or 2 if B is thienyl;

more especially compounds wherein R⁵ denotes (C₁₋₄)alkyl, hydroxy,(C₁₋₄)alkoxy, methanesulphonyloxy or methanesulphonamido, or twoadjacent R⁵ substituents together denote --O--CH₂ --O-- or --O--CH₂--CH₂ --O--;

wherein R⁵ represents hydroxy, (C₁₋₄)alkoxy, methanesulphonyloxy ormethanesulphonamido, or two adjacent R⁵ substituents together represent--O--CH₂ --O-- or --O--CH₂ --CH₂ --O--; particularly

wherein R⁵ represents hydroxy, methoxy, methanesulphonyloxy ormethanesulphonamido, or two adjacent R⁵ substituents together represent--O--CH₂ --O--; particularly compounds wherein R⁵ is methoxy;

wherein q is zero;

wherein B is phenyl and q is two, preferably wherein the two R⁵substituents are in positions 2 and 3.

Mention should also be made of compounds wherein R³ and R⁴ together withthe nitrogen atom to which they are bound represent morpholino,pyrrolidinyl or piperazinyl (which is N-substituted by methoxyphenyl,phenethyl or 2-pyrimidinyl).

Of the above-mentioned groups of compounds the preferred ones are thosewherein

R² represents hydroxy, (C₁₋₄)alkoxy, methanesulphonyloxy ormethanesulphonamido, or two adjacent R² substituents together represent--O--CH₂ --O-- or --O--CH₂ --CH₂ --O--; particularly

wherein R² is methoxy.

B. Of the compounds of formula I wherein R¹ is ##STR42## the compoundswhich may be mentioned as being of particular interest are those wherein

R³ denotes hydrogen; and R⁴ denotes hydrogen, C₃₋₆ -alkenyl; C₃₋₆-alkynyl; C₃₋₆ -cycloalkyl; C₃₋₆ -cycloalkenyl; straight-chained orbranched C₁₋₆ -alkyl, which may optionally be mono- or polysubstitutedwith the following substituents of groups a) to c), which may beidentical or different:

a) halogen; cyano; hydroxy; mercapto; C₁₋₄ -alkoxy; C₁₋₄ -alkylthio;amino; mono-C₁₋₄ -alkylamino; di-C₁₋₄ -alkylamino (wherein the alkylradicals may be identical or different), phenoxy (wherein the phenylgroup may be substituted as in (b)),

b) phenyl; optionally mono- or polysubstituted (with identical ordifferent substituents) by the groups halogen, trifluoromethyl, C₁₋₄-alkoxy, hydroxy, mercapto, C₁₋₄ -alkylthio, C₁₋₄ -alkyl, amino,mono-C₁₋₄ -alkylamino, di-C₁₋₄ -alkylamino (wherein the alkyl groups maybe identical or different), C₂₋₃ -acylamino, C₂₋₃ -acyloxy and the--O--CH₂ --O-- or --O--(CH₂)₂ --O-- group vitally bound to the phenylsystem,

c) a 5- or 6-membered saturated or wholly or partially unsaturatedmonocyclic heterocycle having up to 3 heteroatoms selected from thegroup comprising N, O and S; and as a bicyclic heterocycle indole(whilst the above-mentioned heterocycles may be mono- or polysubstitutedby C₁₋₄ -alkyl), C₃₋₆ -cycloalkyl; C₅ - or C₆ -cycloalkenyl; C₂₋₃ -acyl;C₁₋₄ -alkylsulphonyl; or phenyl (which may in turn be substituted up tothree times as described under b);

or R³ is hydrogen and R⁴ is phenyl which may be substituted as specifiedunder b) above;

R₃ and R₄ independently of each other denote C₁₋₄ -alkyl, which mayoptionally be phenyl-substituted, whilst the phenyl substituent may inturn be substituted as under b) hereinbefore; or

R³ and R⁴ together with the nitrogen atom to which they are bound denotea wholly or partially saturated heterocyclic 5- or 6-membered ring(which may also contain up to two further heteroatoms selected from thegroup N, O, S), whilst the heterocyclic group thus obtained may besubstituted by C₁₋₄ -alkyl, hydroxy, phenyl or benzyl (whilst thisphenyl group or the phenyl group of the benzyl group is substituted asunder b) hereinbefore).

Within the description of general formula I is included in particular3,4-dihydroisoquinoline derivatives wherein NR³ R⁴ is ##STR43## whereinX¹ represents phenyl mono- or disubstituted by trifluoromethyl orethoxy, phenyl substituted by methoxy and fluorine or 2-methoxyphenyl,

X² is --CH₂ --CH₂ -- or --CH₂ --CH(CH₃)--, and

X³ is 2,3,4-trimethoxyphenyl, 2,3-dimethoxyphenyl, 2,6-dimethoxyphenyl,3,6-dimethoxyphenyl, thienyl, phenyl mono- or disubstituted bytrifluoromethyl or ethoxy, or phenyl substituted by methoxy andfluorine,

or the pharmaceutically acceptable salts thereof.

The preferred compounds (I) are those wherein

NR³ R⁴ is ##STR44##

Carbocyclically and heterocyclically anellated dihydropyridines of theformula ##STR45## and the tautomeric forms thereof, wherein R³ denoteshydrogen and

R⁴ denotes hydrogen; straight-chained or branched unsubstituted C₁₋₅-alkyl; allyl; propargyl; C₃₋₆ -cycloalkyl; 3-chlorophenyl;2-methyl-3-chlorophenyl; or C₁₋₃ -alkyl, which is monosubstituted withone of the substituents of groups d) to f) listed below;

d) cyano, hydroxy, methoxy, dimethylamino

e) phenyl, 3,4-methylenedioxyphenyl, phenyl substituted by one, two or 3methoxy groups, 3-hydroxy-4-methoxyphenyl,

f) morpholino, pyridin-2-yl, indol-3-yl, furan-2-yl, thiophen-2-yl,pyridin-3-yl, pyridin-4-yl;

R³ and R⁴ independently of each other denote methyl; ethyl;3-cyanopropyl; benzyl; or 3,4,5-trimethoxyphenethyl or

R³ and R⁴ together with the nitrogen atom to which they are boundrepresent morpholine; thiomorpholine; pyrrolidine; piperazine;4-methylpiperazine; 4-benzylpiperazine or 4-(2-methoxyphenyl)piperazine;and

A represents the anellated ring systems ##STR46## are preferred.

The phenyl group of the compound of general formula I may contain 1, 2or 3 R substituents. These R substituents may be identical or differentfrom one another. Preferably R is C₁₋₄ -alkyl (preferably methyl),halogen (preferably fluorine, chlorine or bromine) or CF₃.

C. Of the compounds of formula I wherein R¹ is unsubstituted phenyl,particular mention may be made of those compounds wherein

R³ is hydrogen and

R⁴ is hydrogen, C₃₋₆ -alkenyl; C₃₋₆ -alkynyl-, C₃₋₆ -cycloalkyl; C₃₋₆-alkyl anellated with a benzo group; C₃₋₆ -cycloalkenyl;straight-chained or branched C₁₋₁₀ -alkyl, which may optionally be mono-or polysubstituted with the following substituents of groups a) to c),which may be identical or different:

a) cyano; hydroxy; C₁₋₄ -alkoxy; C₁₋₄ -alkylthio; C₁₋₄-alkyloxycarbonyl; amino; mono-C₁₋₄ -alkylamino; di-C₁₋₄ -alkylamino(wherein the alkyl radicals may be identical or different), phenoxy(wherein the phenyl group may be substituted as under (b); naphthoxy;

b) phenyl; optionally mono-, di- or trisubstituted (with identical ordifferent substituents) by the groups halogen, trifluoromethyl, C₁₋₄-alkoxy, hydroxy, mercapto, C₁₋₄ -alkylthio, C₁₋₄ -alkyl, azido, amino,mono-C₁₋₄ -alkylamino, di-C₁₋₄ -alkylamino (wherein the alkyl groups maybe identical or different), C₂₋₃ -acylamino, C₂₋₃ -acyloxy and the group--O--CH₂ --O-- or --O--(CH₂)₂ --O-- vicinally bound to the phenylsystem;

c) a 5- or 6-membered saturated or wholly or partially unsaturatedmonocyclic heterocycle having up to 3 heteroatoms from the groupcomprising N, O and S; and as a bicyclic heterocycle indole (whilst theabove-mentioned heterocycles may be mono- or polysubstituted by C₁₋₄-alkyl) C₃₋₁₀ -cycloalkyl (optionally bridged cycloalkyl); C₅ - or C₆-cycloalkenyl; C₂₋₃ -acyl; C₁₋₄ -alkylsulphonyl;

or R⁴ is phenyl which may be substituted as specified under (b)hereinbefore;

R³ and R⁴ independently of each other are each defined as for R⁴ above,and preferably represent C₁₋₆ -alkyl, which may optionally bephenyl-substituted, whilst the phenyl substituent may in turn besubstituted as under b) hereinbefore; or

R³ and R⁴ together with the nitrogen atom to which they are boundrepresent a wholly or partially saturated heterocyclic 5- or 6-memberedring (which may also contain up to 2 further heteroatoms selected fromthe group comprising N, O and S), whilst the heterocyclic group thusobtained may be substituted by C₁₋₄ -alkyl, hydroxy or (CH₂)_(q") R⁸(where q"=0,1,2,3 or 4); and

R⁸ denotes a phenyl radical or phenoxy radical wherein, if desired, thephenyl group is substituted as under b) hereinbefore, or R⁸ denotesnaphthoxy;

A denotes the anellated ring systems ##STR47## wherein R² is ashereinbefore defined, more particularly wherein R³ is hydrogen and

R⁴ is hydrogen; C₃₋₆ -alkenyl; C₃₋₆ -alkynyl; C₃₋₆ -cycloalkyl; C₃₋₆-cycloalkenyl; straight-chained or branched C₁₋₆ -alkyl which mayoptionally be mono- or polysubstituted with the following substituentsof groups a) to c), which may be identical or different:

a) halogen; cyano; hydroxy; mercapto; C₁₋₄ -alkoxy; C₁₋₄ -alkylthio;amino; mono-C₁₋₄ -alkylamino; di-C₁₋₄ -alkylamino (wherein the alkylradicals may be identical or different), phenoxy (wherein the phenylgroup may be substituted as under (b),

b) phenyl; optionally mono- or polysubstituted (with identical ordifferent substituents) by the groups halogen, trifluoromethyl, C₁₋₄-alkoxy, hydroxy, mercapto, C₁₋₄ -alkylthio, C₁₋₄ -alkyl, amino,mono-C₁₋₄ -alkylamino, di-C₁₋₄ -alkylamino (wherein the alkyl groups maybe identical or different), C₂₋₃ -acylamino,-C₂₋₃ -acyloxy and the group--O--CH₂ --O-- or --O--(CH₂)₂ --O-- vicinally bound to the phenyl system

c) a 5- or 6-membered saturated or wholly or partially unsaturatedmonocyclic heterocycle having up to 3 heteroatoms from the group N, O,S; and as a bicyclic heterocycle indole (whilst the above-mentionedheterocycles may be mono- or polysubstituted by C₁₋₄ -alkyl), C₃₋₆-cycloalkyl; C₅ - or C₆ -cycloalkenyl; C₂₋₃ -acyl; C₁₋₄ -alkylsulphonyl;or phenyl (which may in turn be substituted up to three times asdescribed under b);

R³ is hydrogen and

R⁴ is phenyl which may be substituted as specified under (b)hereinbefore;

R³ and R⁴ independently of each other denote C₁₋₄ -alkyl, which mayoptionally be phenyl-substituted, whilst the phenyl substituent may inturn be substituted as under b) hereinbefore; or

R³ and R⁴ together with the nitrogen atom to which they are bound denotea wholly or partially saturated, heterocyclic 5- or 6-membered ring(which may also contain up to 2 further heteroatoms from the group N, O,S), whilst the resulting heterocycle may be substituted by C₁₋₄ -alkyl,hydroxy or (CH₂)_(q") --R⁸ (wherein q"=0 or 1) and

R⁸ denotes a phenyl radical which may optionally be substituted as underb) hereinbefore;

A denotes the anellated ring systems ##STR48## wherein R² is ashereinbefore defined.

General formula I as described covers, in particular,3,4-dihydroisoquinoline derivatives of the general formula ##STR49##wherein NR³ R⁴ represents ##STR50## wherein X¹ denotes phenyl which ismono- or disubstituted by trifluoromethyl or ethoxy, phenyl which issubstituted by methoxy and fluorine, or 2-methoxyphenyl,

X² denotes --CH₂ --CH₂ -- or --CH₂ CH(CH₃)--, and

X³ denotes 2,3,4-trimethoxyphenyl, 2,3-dimethoxyphenyl,2,6-dimethoxyphenyl, 3,6-dimethoxyphenyl, thienyl, phenyl which is mono-or disubstituted by trifluoromethyl or ethoxy, or phenyl substituted bymethoxy and fluorine,

or the pharmaceutically acceptable salts thereof.

Particularly preferred compounds are those wherein

NR³ R⁴ denotes ##STR51## wherein X¹ is 2-methoxyphenyl, which mayadditionally be substituted by fluoro,

X² is --CH₂ --CH₂ -- and

X³ is 2,3,4-trimethoxyphenyl, 2,3-dimethoxyphenyl, 2,6-dimethoxyphenyl,3,6-dimethoxyphenyl, 2- or 3-thienyl, phenyl substituted bytrifluoromethyl or ethoxy, or phenyl substituted by methoxy andfluorine.

Particular mention should be made of compounds of the formula ##STR52##and the tautomeric forms thereof wherein R³ is hydrogen and

R⁴ is hydrogen; straight-chained or branched unsubstituted C₁₋₅ -alkyl;allyl; propargyl; C₃₋₆ -cycloalkyl; 3-chlorophenyl;2-methyl-3-chlorophenyl; or C₁₋₃ -alkyl, which is monosubstituted withone of the substituents of groups d) to f) listed hereinafter;

d) cyano, hydroxy, methoxy, dimethylamino

e) phenyl, 3,4-methylenedioxyphenyl, phenyl substituted by one, two or 3methoxy groups, 3-hydroxy-4-methoxyphenyl,

f) morpholino, pyridin-2-yl, indol-3-yl, furan-2-yl, thiophen-2-yl,pyridin-3-yl, pyridin-4-yl

R³ and R⁴ independently of each other denote methyl; ethyl;3-cyanopropyl; benzyl; or 3,4,5-trimethoxyphenethyl or

R³ and R⁴ together with the nitrogen atom to which they are bound denotemorpholine; thiomorpholine; pyrrolidine; piperazine; 4-methylpiperazine;4-benzylpiperazine; or 4-(2-methoxyphenyl)piperazine; and ##STR53##denotes the anellated ring systems ##STR54## wherein R² is ashereinbefore defined.

The compounds of formula I may be prepared by methods known per se,preferably according to the method described in German PatentApplication P 37 18 570.5, EP 358 957, EP 37 934 and EP 251 794.

In the presence of a condensing agent a malonic acid amide of generalformula IV ##STR55## wherein R¹, R², R³, R⁴ and m are as hereinbeforedefined and Ar denotes phenyl or 2- or 3-thienyl, may be cyclised intothe corresponding compounds.

Suitable condensing agents for this process include strong Lewis acidssuch as phosphorusoxychloride, phosphoruspentachloride,phosphorustrichloride, phosphoruspentoxide, titanium tetrachloride,boron trifluoride, tin tetrachloride, and also inorganic acids such aspolyphosphoric acid, sulphuric acid, fluorosulphonic acid andhydrofluoric acid, or mixtures of condensing agents such as a mixture ofphosphorusoxychloride and phosphoruspentachloride, or a mixture ofphosphoruspentoxide and (C₁₋₄)alkylsulphonic acid, e.g. with a P₂ O₅-content of about 10% by weight.

The cyclisation may be carried out in the presence or absence of asolvent. All inert solvents are suitable provided that they havesufficient solubility for the reactants and a high enough boiling point,e.g. benzene, alkylbenzenes (e.g. toluene, xylene), chlorobenzenes,chloroform, acetonitrile and decalin. According to a preferredalternative embodiment of the process the condensing agent, e.g.phosphorusoxychloride or a (C₁₋₄)alkylsulphonic acid/phosphoruspentoxidemixture is used without the addition of solvents.

Preferably, the cyclisation is carried out using phosphorusoxychlorideor, in difficult cases, with a mixture of phosphoruspentoxide and(C₁₋₄)alkylsulphonic acid (preferably methanesulphonic acid). Thereaction may be carried out within a wide temperature range, preferablywith warming or heating to 50° C. up to about the boiling point of thereaction mixture.

The reaction time required will range from 2 to 15 hours depending onthe starting compound of formula IV.

The compounds of formula I are bases and may be converted in the usualway with organic or inorganic acids and salt-forming agents andcomplexing agents into any desired physiologically acceptable adducts(salts).

Examples of acids suitable for salt formation include hydrochloric acid,hydrobromic acid, hydriodic acid, hydrofluoric acid, sulphuric acid,phosphoric acid, nitric acid, acetic acid, propionic acid, butyric acid,caproic acid, valetic acid, oxalic acid, malonic acid, succinic acid,maleic acid, fumaric acid, lactic acid, tartaric acid, citric acid,malic acid, benzoic acid, p-hydroxybenzoic acid, phthalic acid, cinnamicacid, salicylic acid, ascorbic acid, methanesulphonic acid and the like.

The compounds may be given orally, parenterally or topically. Thedesired therapeutic dose depends on the indication and form ofpreparation and can be determined experimentally. Suitable formsinclude, for example, tablets, capsules, suppositories, solutions,syrups, emulsions, aerosols or dispersible powders. Tablets may beproduced, for example, by mixing the active substance or substances withknown excipients, e.g. inert diluents such as calcium carbonate, calciumphosphate or lactose, disintegrants such as corn starch or alginic acid,binders such as starch or gelatine, lubricants such as magnesiumstearate or talc and/or agents for obtaining delayed release, such ascarboxypolymethylene, carboxymethylcellulose, cellulose acetatephthalate or polyvinylacetate. The tablets may also consist of severallayers.

Coated tablets may be produced analogously by coating cores made in thesame way as the tablets with substances conventionally used for tabletcoatings, e.g. collidone or shellack, gum arabic, talc, titanium dioxideor sugar. In order to obtain delayed release or avoid incompatibilities,the core may also consist of several layers. Similarly, the tabletcoating may consist of several layers to achieve delayed release, whilstthe excipients mentioned for the tablets may be used.

Syrups containing the active substances or combinations of activesubstances according to the invention additionally contain a sweetenersuch as saccharin, cyclamate, glycerol or sugar as well as a flavourenhancer, e.g. a flayouting such as vanillin or orange extract. They mayalso contain suspension adjuvants or thickeners such as sodiumcarboxymethylcellulose, wetting agents, e.g. condensation products offatty alcohols with ethylene oxide or preservatives such asp-hydroxybenzoates.

Injectable solutions are produced in the usual way, e.g. by addingpreservatives such as p-hydroxybenzoates or stabilisers such as alkalimetal salts of ethylene diamine tetraacetic acid, and are thentransferred into injection vials or ampoules.

Capsules containing one or more active substances or combinations ofactive substances may be prepared for example by mixing the activesubstances with inert carriers such as lactose or sorbitol andencapsulating them in gelatine capsules.

Suitable suppositories may be produced for example by mixing withcarriers provided for this purpose, such as neutral fats orpolyethyleneglycol or derivatives thereof.

The compounds may be given both enterally and parenterally. The proposeddose for oral administration is 0.1 to 500 mg of active substance perdose, and for intravenous use 0.05 to 150 mg per dose. The desiredtherapeutic dose depends on the indication and form of preparation andcan be determined experimentally.

The pharmaceutical compositions are suitable for oral or parenteral andpossibly topical use. The pharmaceutical forms are predominantly plainor coated tablets, ampoules and syrups. The individual dose of thesepreparations is between 1.0 and 200 mg, preferably 20 to 50 mg per 75 kgof body weight. Depending on the severity of the case, 1 to 3 singledoses are generally administered per day.

The Examples which follow are intended to illustrate the invention:

EXAMPLE 13,4-Dihydro-1-benzyl-6,7-dimethoxy-α-[di-2-(2,3,4-trimethoxyphenyl)ethyl]aminocarbonyl-isoquinolinehydrochloride##STR56##

a) 2-(3,4-Dimethoxyphenyl)ethylaminocarbonyl-phenylaceticacid-N,N-di-[2-(2,3,4-trimethoxyphenyl)ethyl]amide

To a solution of 18.0 g (52.4 mmol) ofmonoethylphenylmalonate-2-(3,4-dimethoxyphenyl)ethylamide in 150 ml ofanhydrous dimethylformamide are added, at ambient temperature 9.0 g(55.5 mmol) of N,N'-carbonyldiimidazole in batches. After 30 minutes18.0 g (44.3 mmol) of di-[2-(2,3,4-trimethoxyphenyl)ethyl]amine areadded and the mixture is stirred for 30 minutes. Then the solvent isdistilled off in vacuo, the residue is taken up in 1.5 liters of CH₂ Cl₂and extracted twice with 250 ml of water and 200 ml of 1N HCl, one afterthe other. The organic phase is evaporated down after drying over Na₂SO₄ and, after purification over a silica gel column (eluant: CH₂ Cl₂/MeOH 100:2) the residue is crystallised from ethyl acetate/ether.Yield: 35.5 g

b) 35.0 g (47.5 mmol) of amide (from step a) and 15 ml (164 mmol) ofphosphorusoxychloride are heated to boiling for 30 minutes in 150 ml ofanhydrous acetonitrile. After the reaction has ended (monitored by thinlayer chromatography) the solvent and any unused phosphorusoxychlorideare distilled off in vacuo. The residue is mixed with ice water, madealkaline with soda solution and extracted in batches with about 1 literof CH₂ Cl₂. The organic phase is washed with water, dried over Na₂ SO₄and evaporated down. The residue is purified twice over a silica gelcolumn (1st eluant: CH₂ Cl₂ :MeOH 100:2→100:4 rising; 2nd eluant: CH₂Cl₂ /ethyl acetate 1:1).

The hydrochloride is formed from the purified product (6.5 g) bydissolving in about 50 ml of ethanol and adding alcoholic hydrochloricacid. After evaporation and drying under a high vacuum at 50° C. 11.5 gof the desired product remain. (M.p.56°-64° C., amorphous)

EXAMPLE 2 2-Phenylmalonic acid-N-(2-(3,4-dimethoxyphenyl)ethyl)N',N'-di-(2-(2-fluorophenyl)ethyl)-diamide

To a solution of 17.2 g (0.05 mol) of phenylmalonic acidN-(2-(3,4-dimethoxyphenyl)ethyl)-monoamide in 200 ml of anhydrous CH₂Cl₂, 8.1 g (0.05 mol) of N,N'-carbonyldiimidazole are added in batchesat ambient temperature and with stirring. After about 30 minutes asuspension consisting of 14.9 g (0.05 mol) ofdi-(2-(2-fluorophenyl)ethyl)amine-hydrochloride and 5.1 g=6.4 ml (0.05mol) of triethylamine in 100 ml of anhydrous CH₂ Cl₂ is stirred into thereaction mixture. After 15 hours' stirring, 200 ml of water are added,the mixture is acidified with dilute hydrochloric acid, the organicphase is separated off and the aqueous phase is extracted 3 times with100 ml of CH₂ Cl₂. The combined organic phases are dried over Na₂ SO₄and evaporated down.

R,S (3,4-Dihydro-6,7-dimethoxyisoquinolin-1-yl)-2-phenylN,N-di-(2-(2-fluorophenyl)ethyl-acetamide-oxalate

A mixture of 29.8 g (0.049 mol) of amide, 150 ml of anhydrous CH₂ Cl₂and 17.5 g=9.4 ml (0.10 mol) of POCl₃ is refluxed for 10 hours. Afterthe reaction has ended (TLC monitoring) the mixture is stirred into acombination of 400 ml of ice water and 200 ml of CH₂ Cl₂. Then it isneutralised with a saturated soda solution, the organic phase isseparated off, the aqueous phase is extracted 3 times with 100 ml of CH₂Cl₂. The combined phases are dried over Na₂ SO₄ and evaporated down invacuo. The residue is purified over silicic acid (eluant: CH₂ Cl₂=methanol=100:2) and then dissolved in a little ethanol and convertedinto the oxalate by adding a stoichiometric quantity of an alcoholicoxalic acid solution. The oxalate is also crystallised by the additionof ether. M.p.: 144°-146° C.

EXAMPLE 3(R,S)-(3,4-Dihydro-6,7-dimethoxyisoquinolin-1-yl)-2-(4-methoxyphenyl)-N,N-di(2-(2,3,4-trimethoxyphenyl)ethyl)acetamide-oxalate

Ingredients:

5 g (6.57 mMol) of 4-methoxyphenylmalonicacid-N-(2-(3,4-dimethoxyphenyl)ethyl)-N'-(2-(2,3,4-trimethoxyphenyl)-ethyl-diamide("diamide") 18 ml of acetonitrile, 3.02 g (19.7 mMol) ofphosphorusoxychloride, 600 mg (6.66 mMol) of oxalic acid, anhydrous, 200ml of ether

Method:

The reaction mixture of the "diamide", acetonitrile andphosphorusoxychloride is refluxed for 1 hour under protective N₂. Aftercooling with ice water it is diluted with 100 ml of ethyl acetate andthen washed twice each with ice water, 50 ml of saturated NaHCO₃solution, water and saturated NaCl solution. The organic phase is driedover MgSO₄ and poured, with stirring, into a solution of 706 mg (7.84mMol) of anhydrous oxalic acid in 200 ml of absolute ether.

The reaction product was initially precipitated as an oil as the oxalicacid salt, which crystallised after standing for some time withstirring. After standing overnight in the refrigerator it was suctionfiltered, washed with ether and dried. M.p.: 107°-110° C. The structurewas confirmed by NMR spectroscopy. R_(f) values: 0.53 (ethyl acetate)R_(f) values: 0.6 (acetonitrile:H₂ O=9:1)

Preparation of the starting compound:

Part A Diethyl 4-methoxyphenylmalonate

Ingredients:

150 ml of absolute ethanol

7.5 g (0.33 mol) of sodium

300 ml of diethylcarbonate

62.5 g (0.3 mol) of ethyl 4-methoxyphenylacetate

Method:

Sodium is dissolved in absolute ethanol and evaporated to dryness invacuo. Whilst cooling with ice water the residue is combined withdiethylcarbonate and ethyl 3-chlorophenylacetate, with stirring. Thenthe ethanol is slowly (2-3 hours) distilled off through a 40 cm column(Raschig rings) under a high vacuum at 40°-70° C. and 200 mM. Aftercooling, the mixture is acidified with 30 ml of glacial acetic acid andmixed with 150 ml of water. The oil precipitated is washed successivelywith water and saturated NaCl solution and dried over MgSO₄. The residueis distilled in a bulb tube using an oil pump; b.p.: 0.5 mM: 150°-155°C.

Monoethyl 4-methoxyphenylmalonate

Ingredients:

52.2 g (0.196 mol) of diethylester

120 ml of ethanol

120 ml of water

12.3 g (0.22 mol) of KOH in 60 ml of water and 60 ml of ethanol

Method:

An ethanolic solution of diethyl 4-methoxyphenylmalonate is mixed withthe aqueous alcoholic KOH solution, with stirring and cooling with ice,and stirred for 75 minutes. Then it is acidified with a saturated citricacid solution and extracted 3 times with methylene chloride. The organicphase is washed successively with water and saturated NaCl solution,dried over MgSO₄ and the solvent is distilled off in vacuo. Thecrystalline residue is recrystallised from methylene chloride/petroleumether (40-800).

Yield: 34.4 g (73.6% of theory).

The structure was confirmed by NMR spectroscopy.

Monoethyl N-[2,3,4-dimethoxyphenyl)ethyl)-4-methoxyphenyl-malonate-amide

Ingredients:

34.4 g (0.144 mol) of monoethyl 4-methoxy-phenylmalonate

150 ml of anhydrous tetrahydrofuran

23.3 g (0.144 mol) of N,N'-carbonyldiimidazole

26.1 g (0.144 mol) of 2-(3,4-dimethoxyphenyl)ethylamine in 50 ml ofanhydrous tetrahydrofuran

Method:

The carbonyldiimidazole is added in batches at 50° C., with stirring, tothe solution of the 4-methoxyphenylmalonic acid hemiester in THF. After30 minutes' stirring at ambient temperature the amine is added whilstcooling with ice and the mixture is stirred for 16 hours at ambienttemperature. The reaction mixture is evaporated down and the residue istaken up in CH₂ Cl₂. It is washed twice with water, then 10% KHSO₄solution, saturated NaHCO₃ solution, water and saturated salinesolution. After drying over MgS₄ the organic phase is evaporated downand the oily residue (7.7 g) is crystallised from ethyl acetate.

4-Methoxy-phenylmalonic acid-N-(2(3,4-dimethoxyphenyl)ethyl)-amide

Ingredients:

44.16 g (0.11 mol) (4-methoxy-phenylmalonic acid hemiester amide)

300 ml methanol

120 ml (0.12 mol) 1N sodium hydroxide solution

Method:

At 5°-10° C. the sodium hydroxide solution is stirred into themethanolic solution of the hemiester amide for 30 minutes. After 3hours' stirring at ambient temperature it is evaporated down, dilutedwith water, then extracted twice with CH₃ Cl. The aqueous phase isadjusted to pH 1 with concentrated HCl and extracted twice with CH₃ Cl.After washing with saturated saline solution the organic phase is driedover MgSO₄ and evaporated down. The crystalline residue is crystallisedfrom CH₃ Cl.

Part B 2-(2,3,4-Trimethoxyphenyl)-1-nitro-ethers

Ingredients:

400 g (2.04 mol) of 2,3,4-trimethoxybenzaldehyde

1740 g of glacial acetic acid

172.6 g (2.24 mol) of anhydrous ammonium acetate

656 ml of nitromethane

Method:

The mixture of benzaldehyde, ammonium acetate, nitromethane and glacialacetic acid is stirred for 30 minutes at boiling temperature undernitrogen protection. The mixture is cooled to -5° C. and poured into 5liters of ice water with stirring. The viscous residue is extractedexhaustively with CH₂ Cl₂. The organic phase is washed with water, driedover MgSO₄ and evaporated down. The brown oily residue (510 g)crystallises on standing. It is dissolved in 470 ml of ethyl acetate andcrystallised by the addition of 3.9 liters of cyclohexane.

2-(2,3,4-Trimethoxyphenyl)ethylamine-hydrochloride

Ingredients:

144.5 g (0.60 mol) "Nitrostyrol"

1.9 l of water

232 ml of conc. hydrochloric acid (analytical grade)

88 g PdC (10%)

Method:

The above reaction mixture is hydrogenated at 60° C. under 35 bar for2.25 hours. Then it is evaporated down in vacuo, the residue is taken upin ethanol and evaporated to dryness. It is then dissolved in ethanoland the reaction product is crystallised by the addition of ether.

2,3,4-Trimethoxybenzylalcohol

Ingredients:

500 g (2.55 mol) 2,3,4-trimethoxybenzaldehyde

5.0 l of methanol 13.0 g of PtO₂

Method:

The reduction is carried out at 20° C. under 5 bar and is complete after30 minutes. After the solvent has been evaporated off the residue (501.5g) is distilled in a high vacuum (B.p. 116°; 0.5 mbar).

2,3,4-Trimethoxybenzylchloride

Ingredients:

50.0 g (0.25 mol) 2,3,4-trimethoxybenzylalcohol

800 ml of anhydrous methylene chloride

59.4 g (0.5 mol) of thionylchloride

Method:

The solution of the alcohol in anhydrous CH₂ Cl₂ is slowly mixed withSOCl₂ with stirring and cooling with ice and common salt. The mixture isstirred for a further 15 minutes in the cold, then for 2 hours atambient temperature. The solvent and excess thionylchloride areeliminated in vacuo, the residue is taken up in CH₂ Cl₂ and shakensuccessively with saturated NaHCO₃ solution, water and saturated salinesolution. After drying over MgSO₄ the solvent is removed in vacuo andthe residue is distilled in a bulb tube furnace using an oil pump (b.p.118° C.; 0.1 mbar).

2,3,4-Trimethoxybenzylcyanide

Ingredients:

75.8 g (0.35 mol) 2,3,4-trimethoxybenzylchloride

700 ml of anhydrous acetone

3.45 g (0.023 mol) of NaI

25.7 g (0.53 mol) of dried powdered NaCN

Method:

The reaction mixture consisting of benzyl chloride, NaI and NaCN inanhydrous acetone is stirred for 20 hours at boiling temperature. Aftercooling it is suction filtered and the solvent is eliminated in vacuo.The residue is dissolved in ethyl acetate, shaken first with water thenwith saturated saline solution and dried over MgSO₄. After removal ofthe solvent the residue is distilled in a bulb tube furnace at 0.015mbar (b.p. 135° C.).

2,3,4-Trimethoxyphenylacetic acid

Ingredients:

138.5 g (0.67 mol) 2,3,4-trimethoxybenzylcyanide,

53.5 g (1.34 mol) NaOH dissolved in 215 ml of water

Method:

The mixture of benzyl cyanide and aqueous sodium hydroxide solution isrefluxed for 7 hours, then after cooling acidified with 6N H₂ SO₄ andextracted three times with CH₂ Cl₂. The organic phase is washed withwater and saturated NaCl solution and dried over MgSO₄. After removal ofthe solvent the residue is dissolved in 200 ml of CH₂ Cl₂ andcrystallised by the addition of 1500 ml of cyclohexane.

2,3,4-Trimethoxyphenyl-N-(2-(2,3,4-trimethoxyphenyl)ethyl)-acetamide

Ingredients:

72.4 g (0.32 mol) 2,3,4-trimethoxyphenylacetic acid

400 ml anhydrous tetrahydrofuran

51.8 g (0.32 mol) N,N'-carbonyldiimidazole (CDI)

79.3 g (0.32 mol) 2-(2,3,4-trimethoxyphenyl)ethylamine-hydrochloride

500 ml anhydrous tetrahydrofuran

32.4 ml (0.32 mol) triethylamine

Method:

The phenylacetic acid dissolved in anhydrous THF is converted into theimidazolide with stirring at 5° C. by the batch-wise addition of CDI.After 30 minutes a suspension of amine hydrochloride, triethylamine inanhydrous THF is stirred in at ambient temperature. After 16 hours themixture is evaporated down in vacuo and the residue is divided betweenCH₂ Cl₂ and 2N HCl. Then it is washed successively with water, saturatedNaHCO₃ solution, water and saturated NaCl solution. After drying overMgSO₄ the organic phase is evaporated down, the residue is dissolved in200 ml of ethyl acetate and the product is crystallised by the additionof 700 ml of cyclohexaneo

Di-2-(2,3,4-trimethoxyphenyl)ethylamine-hydrochloride

Ingredients:

113.4 g (0.27 mol) "phenylacetic acid amide"

470 ml anhydrous tetrahydrofuran

270 ml (0.54 mol) BH₃.S(CH₃)₂ in THF (2 mol/l)

Method:

At 65° C. the borane complex mixture is added dropwise to the solutionof the acid amide in anhydrous THF under protective N₂. After theaddition has ended the mixture is stirred for a further 15 minutes at65° C. Then the reaction mixture is cooled to 5° C. It is carefullyacidified with methanolic hydrochloric acid, evaporated down in vacuo(drawn off) and the residue is crystallised from ethanol with theaddition of ether.

Part C 4-Methoxy-phenylmalonicacid-N-(2-(3,4-dimethoxyphenyl)ethyl)-N'-(2-(2,3,4-trimethoxyphenyl)ethyl)-di-amide

Ingredients:

3.73 g (10 mMol) 4-methoxy-phenylmalonic acid hemiamide

25 ml anhydrous THF

1.62 g (10 mMol) N,N'-carbonyldiimidazole

4.1 g (10 mMol) di-2-(2,3,4-trimethoxyphenyl)ethylaminehydrochloride

40 ml anhydrous THF

1.01 g (10 mMol) triethylamine=1.39 ml

Method:

The carbonyldiimidazole is added in batches to the solution of themonoamide in THF at 5° C. with stirring. The reaction mixture is stirredfor 30 minutes at ambient temperature. Then, whilst cooling with ice, itis reacted with a suspension of the amine-hydrochloride in THF andtriethylamine. After 16 hours' stirring at ambient temperature it isevaporated down and the residue is dissolved in ethyl acetate. Theorganic phase is washed successively with water, 5% KHSO₄ solution,saturated NaHCO₃ solution, water and unsaturated NaCl solution. Afterdrying over MgSO₄ the mixture is evaporated down, the residue (7.2 g) ispurified on 210 g of silica gel (eluant: ethyl acetate/n-hexane=2:1).

The following Tables contain examples of compounds according to theinvention, whilst Tables 10 to 20 list new compounds.

                  TABLE 1                                                         ______________________________________                                        Structural type:                                                               ##STR57##                                                                                                         Salt                                     No.  X                      Structure                                                                              form                                     ______________________________________                                         1.  OCH.sub.3              II       --                                        2.  OC.sub.2 H.sub.5       II       --                                        3.  NHCH.sub.3             I        Cl                                        4.  NHC.sub.2 H.sub.5      I        --                                        5.  NH(CH.sub.2).sub.2 CH.sub.3                                                                          I        Cl                                        6.  NH(CH.sub.2).sub.3 CH.sub.3                                                                          I        Cl                                        7.  NH(CH.sub.2).sub.4 CH.sub.3                                                                          I        Cl                                        8.  NHCH(CH.sub.3).sub.2   I        Cl                                        9.  NHCH.sub.2 CH(CH.sub.3).sub.2                                                                        I        Cl                                       10.  NH(CH.sub.2).sub.2 CH(CH.sub.3).sub.2                                                                I        Cl                                       11.  NHC(CH.sub.3).sub.3    I        Cl                                       12.  NHCH(CH.sub.3)C.sub.2 H.sub.5                                                                        I        Cl                                       13.  NHCH.sub.2 CHCH.sub.2  I        Cl                                       14.  NHCH.sub.2 CCH         I        Cl                                       15.  NH(CH.sub.2).sub.2 OH  II       Cl                                       16.  NHCH.sub.2 CH(OH)CH.sub.3                                                                            I        Cl                                       17.  NH(CH.sub.2).sub.2 OCH.sub.3                                                                         II       Cl                                       18.  NH(CH.sub.2).sub.3 OCH.sub.3                                                                         II       Cl                                       19.  NH(CH.sub.2).sub.2 N (CH.sub.3).sub.2                                                                I        Cl.sub.2                                 20.  NH(CH.sub.2).sub.3 N (CH.sub.3).sub.2                                                                II       Cl.sub.2                                       ##STR58##             II       --                                             ##STR59##             I        Cl                                             ##STR60##             I        --                                             ##STR61##             I        --                                             ##STR62##             I        --                                             ##STR63##             II       --                                             ##STR64##             II       Cl                                             ##STR65##             I        --                                             ##STR66##             I        --                                       30.                                                                                 ##STR67##             II       --                                             ##STR68##             I        --                                       32                                                                                  ##STR69##             II       --                                       33                                                                                  ##STR70##             I        Cl                                             ##STR71##             I        Cl                                             ##STR72##             II       --                                       36.  N(CH.sub.3).sub.2      I        Cl                                       37.  N(C.sub.2 H.sub.5).sub.2                                                                             I        Cl                                             ##STR73##             I        Cl                                       38a.                                                                                ##STR74##             I        Cl                                             ##STR75##             I        Cl                                       40.                                                                                 ##STR76##             I        --                                             ##STR77##             II       --                                             ##STR78##             II       Cl.sub.2                                       ##STR79##             I        --                                             ##STR80##             I        Cl                                       ______________________________________                                    

                  TABLE 2                                                         ______________________________________                                        Structural type:                                                               ##STR81##                                                                    No.  X                     Structure                                                                              Salt form                                 ______________________________________                                        45.  OC.sub.2 H.sub.5      II       --                                              ##STR82##            II       Cl                                              ##STR83##            II       --                                              ##STR84##            II       --                                              ##STR85##            II       --                                        50.                                                                                 ##STR86##            I        --                                        ______________________________________                                    

                  TABLE 3                                                         ______________________________________                                        Structural type:                                                               ##STR87##                                                                    No.  X                     Structure                                                                              Salt form                                 ______________________________________                                        51.  OC.sub.2 H.sub.5      I        --                                                                   II                                                 52.  NH(CH.sub.2).sub.3 CH.sub.3                                                                         I        --                                        53.  NH(CH.sub.2).sub.4 CH.sub.3                                                                         I        --                                        54.  NHCH(CH.sub.3).sub.2  I        Cl                                        55.  NHCH.sub.2 CH(CH.sub.3).sub.2                                                                       I        --                                        56.  NHCH.sub.2 CHCH.sub.2 I        --                                              ##STR88##            I        Cl                                              ##STR89##            I        Cl                                              ##STR90##            I                                                  60.                                                                                 ##STR91##            I                                                        ##STR92##            I        --                                              ##STR93##            I        --                                              ##STR94##            I        Cl                                              ##STR95##            I        --                                              ##STR96##            I        Cl                                              ##STR97##            I        Cl                                              ##STR98##            I        Cl                                              ##STR99##            I        Cl                                        ______________________________________                                    

                  TABLE 4                                                         ______________________________________                                        Structural type:                                                               ##STR100##                                                                   No.     X        Structure      Salt form                                     ______________________________________                                        69.     N(C.sub.2 H.sub.5).sub.2                                                               I                                                            ______________________________________                                    

                  TABLE 5                                                         ______________________________________                                        Structural type:                                                               ##STR101##                                                                   No.   X                  Structure Salt form                                  ______________________________________                                        70.   NHCH.sub.5         I                                                    71.   NHC.sub.2 H.sub.5  I                                                           ##STR102##        I                                                           ##STR103##        I                                                    74.   N(CH.sub.3)C.sub.2 H.sub.5                                                                       I                                                    ______________________________________                                    

                  TABLE 6                                                         ______________________________________                                        Structural type:                                                               ##STR104##                                                                   No.    X                    Structure                                         ______________________________________                                                ##STR105##          I                                                         ##STR106##          I                                                 77.    N(CH.sub.3)C.sub.2 H.sub.5                                                                         I                                                         ##STR107##          II                                                        ##STR108##          I                                                 ______________________________________                                    

                  TABLE 7                                                         ______________________________________                                        Structural type:                                                               ##STR109##                                                                   No.  X                     Structure                                                                              Salt form                                 ______________________________________                                        82.  OC.sub.2 H.sub.5      I                                                                             II       --                                              ##STR110##           II       --                                              ##STR111##           II       --                                              ##STR112##           I II      --                                       86                                                                                  ##STR113##           I        --                                        87.  NHCH.sub.2 CH(CH.sub.3).sub.2                                                                       II       --                                        88.  NH(CH.sub.2).sub.3 N(CH.sub.3).sub.2                                                                II       --                                              ##STR114##           II       --                                        ______________________________________                                    

                  TABLE 8                                                         ______________________________________                                        Structural type:                                                               ##STR115##                                                                   No.    X                Structure  Salt form                                  ______________________________________                                        90.    OC.sub.2 H.sub.5                                                               ##STR116##      II         --                                         ______________________________________                                    

                                      TABLE 9                                     __________________________________________________________________________     ##STR117##                                                                   Structural type I                                                              ##STR118##                                                                   Structural type II                                                            X                            Structural type                                                                       Mp [°C.]                          __________________________________________________________________________    Compound                                                                             ##STR119##            I       56-64                                    B                                                                                    ##STR120##            I       176-184                                  Compound                                                                      (Salt form)                                                                   C                                                                                    ##STR121##            I       166-168                                  D                                                                                    ##STR122##            II      102-104                                  E (Cl)                                                                               ##STR123##            I       187                                      F (--)                                                                               ##STR124##            I       94-96                                    G (Cl)                                                                               ##STR125##            II      139-142                                  H (--)                                                                               ##STR126##            II      133-135                                  J (--)                                                                               ##STR127##            II      143-145                                  K (--)                                                                               ##STR128##            II      96-98                                    L (--)                                                                               ##STR129##            II      118-120                                  M (--)                                                                               ##STR130##            II      112-114                                  N (Cl)                                                                               ##STR131##            I       95-99                                    O (--)                                                                               ##STR132##            I       114-116                                  P (--)                                                                               ##STR133##            I       66-73                                    Q (Cl)                                                                               ##STR134##            II      205-209                                  __________________________________________________________________________

                                      TABLE 10                                    __________________________________________________________________________    Structural type:                                                               ##STR135##                                                                   No.                                                                              X                       Structure                                                                          Salt form                                                                           Mp [°C.]                         __________________________________________________________________________    101                                                                               ##STR136##             I    Oxalate                                                                             138-140                                 102                                                                               ##STR137##             I    Oxalate                                                                             153-155                                 103                                                                               ##STR138##             I    Oxalate                                                                             117-123                                 104                                                                               ##STR139##             I    Oxalate                                                                             154-155                                 105                                                                               ##STR140##             I    Oxalate                                                                             127-129                                 106                                                                               ##STR141##             I    Oxalate                                                                             132-135                                 107                                                                               ##STR142##             I    Oxalate                                                                             145-147                                 108                                                                               ##STR143##             I    Base  118-120                                 109                                                                               ##STR144##             II   Base  118-120                                 110                                                                               ##STR145##             I    Oxalate                                                                             153-156                                 111                                                                              N[(CH.sub.2).sub.5 CH.sub.3 ].sub.2                                                                   I    Oxalate                                                                              81-84                                  112                                                                              N(CH.sub.2 CH.sub.2 CH.sub.3).sub.2                                                                   I    Oxalate                                                                              57-59                                  113                                                                              N[(CH.sub.2).sub.4 CH.sub.3 ].sub.2                                                                   I    Oxalate                                                                             --                                      114                                                                               ##STR146##             I    Oxalate                                                                             125-126                                 115                                                                               ##STR147##             I    Base  103-105                                 116                                                                              N[(CH.sub.2).sub.3 CH.sub.3 ].sub.2                                                                   I    Oxalate                                                                             119-121                                 117                                                                              N[CH.sub.2 CH(CH.sub.3).sub.2 ].sub.2                                                                 I    Oxalate                                                                             130-131                                 118                                                                               ##STR148##             I    Oxalate                                                                             176-178                                 119                                                                               ##STR149##             I    Oxalate                                                                             160-162 (Decomp)                        120                                                                               ##STR150##             I    Oxalate                                                                             170-172                                 121                                                                               ##STR151##             I    Oxalate                                                                             158-160 (Decomp)                        122                                                                               ##STR152##             I    Oxalate                                                                             166-168                                 123                                                                              NH(CH.sub.2).sub.9 CH.sub.3                                                                           I    Oxalate                                                                              99-101                                 124                                                                               ##STR153##             I    Oxalate                                                                             127-143                                 125                                                                               ##STR154##             I    Oxalate                                                                             144-149                                 126                                                                               ##STR155##             I    Oxalate                                                                             137-139                                 127                                                                               ##STR156##             I    HCl   185-187                                 128                                                                              N(CH.sub.2 CN).sub.2    I    Base                                          129                                                                               ##STR157##             II   Base                                          130                                                                               ##STR158##             II   HCl                                           131                                                                               ##STR159##             II   HCl                                           132                                                                              NHCH.sub.2 -Adam(1)     II   HCl                                           133                                                                               ##STR160##             I    Oxalate                                       134                                                                               ##STR161##             I    Base                                          135                                                                               ##STR162##             I    HCl                                           136                                                                               ##STR163##             I    HCl                                           137                                                                               ##STR164##             I    (Oxalate).sub.2                               138                                                                               ##STR165##             I    HCl                                           139                                                                               ##STR166##             I    HCl                                           140                                                                               ##STR167##             I    Base                                          141                                                                              OCH.sub.2 C(CH.sub.3).sub.3                                                                           II   Base                                          142                                                                               ##STR168##             II   Base                                          143                                                                              OCH.sub.2 CH.sub.2 -Adam(1)                                                                           II   HCl                                           144                                                                              OCH.sub.2 CH.sub.2 C(CH.sub.3).sub.3                                                                  II   HCl                                           145                                                                              OCH.sub.2 -Adam(1)      II   Base                                          146                                                                               ##STR169##             II   HCl                                           147                                                                               ##STR170##             II   HCl                                           148                                                                               ##STR171##             I    Oxalate                                                                             118-120                                 149                                                                               ##STR172##             I    HCl   195-197                                 150                                                                               ##STR173##             I    HCl   140-143                                 151                                                                               ##STR174##             I    HCl   193-195                                 152                                                                               ##STR175##             I    HCl   155-158                                 153                                                                               ##STR176##             I    HCl   168-170                                 154                                                                               ##STR177##             I    HCl   172-174                                 __________________________________________________________________________     Adam = Adamantyl                                                         

                                      TABLE 11                                    __________________________________________________________________________     ##STR178##                                                                   R.sub.1   X                   Salt form                                       __________________________________________________________________________     ##STR179##                                                                              ##STR180##         HCl  Mp. 116-127° C.                      ##STR181##                                                                              ##STR182##         BS   Rf: 0.39 (ethyl acetate)                   __________________________________________________________________________

                  TABLE 12                                                        ______________________________________                                         ##STR183##                                                                   No.   R            Structure Salt form                                                                             Mp [°C.]                          ______________________________________                                        1     4-CH.sub.3   I         Oxalate                                          2     4-F          I         Oxalate                                          3     3-Cl         I         Oxalate                                          4     4-Br         I         Oxalate                                          5     2-OCH.sub.3  I         Oxalate                                          6     3-OCH.sub.3  I         Oxalate                                          7     4-OCH.sub.3  I         Oxalate                                          8     3,4-di-OCH.sub.3                                                                           I         Oxalate                                          9     3,4,5-tri-OCH.sub.3                                                                        I         Oxalate                                          10    4-N.sub.3    I         Oxalate                                          11    3-I,4-N.sub.3                                                                              I         Oxalate                                          12    2-CH.sub.3   I         Oxalate                                          13    2-Br         I         Base                                             14    2,4-di-Cl    I         Oxalate                                          15    3-F          I         Oxalate                                          16    2-Cl         I         Base                                             ______________________________________                                    

                                      TABLE 13                                    __________________________________________________________________________     ##STR184##                                                                   No.                                                                              NR.sup.3 R.sup.4      Salt form                                                                          Structure                                                                          Mp (°C.)                                                                    % Inhib.                              __________________________________________________________________________        ##STR185##           1.5 Fu                                                                             I    177-178                                                                            40.44                                    "                     BS   II   158-159                                                                            0.0                                       ##STR186##           BS   II   165-167                                                                            50.64                                     ##STR187##           MS   I    132-133                                                                            67.73                                     ##STR188##           BS   II   84-55                                                                              65.50                                     ##STR189##           MS   I    151-153                                                                            80.89                                    "                     BS   II   110-111                                                                            39.47                                     ##STR190##           MS   I    118-122                                                                            24.03                                     ##STR191##           MS   I    150-154                                                                            43.49                                     ##STR192##           Cl   I    144-147                                                                            15.26                                     ##STR193##           BS   II   131-133                                                                            26.54                                     ##STR194##           Cl   I    114-115                                                                            0.0                                       ##STR195##           BS   II   144-147                                                                            67.75                                 __________________________________________________________________________

                                      TABLE 14                                    __________________________________________________________________________     ##STR196##                                                                   No.                                                                              NR.sup.3 R.sup.4         Salt form                                                                          Structure                                                                          Mp (°C.)                                                                      % Inhib.                         __________________________________________________________________________        ##STR197##              Cl   I    amorphous.sup.x)                                                                     79.17                                ##STR198##              Cl   I    amorphous.sup.x)                                                                     73.55                                ##STR199##              Cl   I    amorphous.sup.x)                                                                     60.82                                ##STR200##              Cl   I + II                                                                             amorphous.sup.x)                                                                     82.76                                ##STR201##              Cl   I    amorphous.sup.x)                                                                     50.90                                ##STR202##              Ox   I    amorphous.sup.x)                                                                     56.20                                ##STR203##              Ox   I    142-144                                     ##STR204##              BS   I    105-106                                                                              22.39                                ##STR205##              Cl.sub.2                                                                           I    208-210                                     ##STR206##              MS   I    amorphous.sup.x)                                                                     79.60                                ##STR207##              Cl   I    amorphous.sup.x)                                                                     36.71                                ##STR208##              Cl   I    amorphous.sup.x)                                                                     30.87                            __________________________________________________________________________     .sup.x) The structure of the substance is characterised by NMR                spectroscopy                                                             

                                      TABLE 15                                    __________________________________________________________________________     ##STR209##                                                                   No.                                                                              NR.sup.3 R.sup.4  Salt form                                                                          Structure                                                                          Mp (°C.)                                                                    % Inhib.                                  __________________________________________________________________________     ##STR210##          Cl   I    134-136                                         ##STR211##          Cl   I    166-168                                         ##STR212##          Cl   I    142-144                                         ##STR213##          BS   I    119  84.43                                     __________________________________________________________________________

                                      TABLE 16                                    __________________________________________________________________________     ##STR214##                                                                      Salt                                                                       No.                                                                              form                                                                             R.sup.a     R.sup.b    NR.sup.3 R.sup.4     Mp.  %                                                                                IC.sub.50           __________________________________________________________________________    MS    HO          CH.sub.3 O                                                                                ##STR215##               87 3.47 ×                                                                  10.sup.-6           MS    CH.sub.3 O  HO         "                         0  --                  MS    CH.sub.3 O  CH.sub.3 O                                                                                ##STR216##                                      OX    CH.sub.3 O  H                                                                                         ##STR217##          60-70                                                                              30.41                  MS                                                                                   ##STR218## H          "                    182-185                                                                            14.65                  OX    HO          H          "                    120-130                     OX    CH.sub.3 O  CH.sub.3   "                    143  10.47                  OX                                                                                   ##STR219## CH.sub.3   "                    156-157                                                                            49.03                  OX    HO          CH.sub.3   "                    80-95                       OX    CH.sub.3    Cl         "                    143-145                                                                            44.65                  OX                                                                                   ##STR220## Cl         "                    130-132                                                                            32.90                  OX    HO          Cl         "                    182-183                     MS                                                                                   ##STR221## CH.sub.3   "                         31.8                   MS    CH.sub.3 O                                                                                 ##STR222##                                                                              "                         22.9                   OX    CH.sub.3 O  CH.sub.3                                                                                  ##STR223##          145-151                                                                            58.3                   OX    CH.sub.3 O  H          "                    115-125                     OX    CH.sub.3 O  Cl         "                    131-135                     OX                                                                                   ##STR224## Cl         "                    135-142                     OX                                                                                   ##STR225## CH.sub.3   "                    162-164                     OX                                                                                   ##STR226## H          "                    145-147                     OX    HO          Cl         "                    187-190                     OX    HO          CH.sub.3   "                    193-200                     OX    HO          H          "                    154-157                     __________________________________________________________________________

                                      TABLE 17                                    __________________________________________________________________________     ##STR227##                                                                   No.                                                                              NR.sup.3 R.sup.4        Salt form                                                                          Mp (°C.)                                                                      % Hem.                                                                             IC.sub.50                         __________________________________________________________________________     ##STR228##                OX   amorphous*                                                                           68.2                                    ##STR229##                Cl   amorphous*                                                                           36.95                                   ##STR230##                Cl          69.7                                    ##STR231##                Cl   amorphous*                                     ##STR232##                BS   amorphous*                                     ##STR233##                Cl   amorphous*                                    HNCH.sub.2 -1-Adam.        Cl   amorphous*                                                                           65.1                                    ##STR234##                BS   amorphous*                                     ##STR235##                Cl   amorphous*                                                                           12.82                                   ##STR236##                BS   amorphous*                                                                           59                                      ##STR237##                Cl   amorphous*                                                                           72.18                                   ##STR238##                Cl   amorphous*                                     ##STR239##                OX-2 amorphous*                                                                           41.06                                   ##STR240##                BS   amorphous*                                                                           7.77                                    ##STR241##                BS   amorphous*                                                                           45.26                                   ##STR242##                OX   amorphous*                                                                           33.06                                   ##STR243##                Cl   199-201                                                                              81.60                                   ##STR244##                Cl   amorphous*                                                                           52.51                                   ##STR245##                Cl   amorphous*                                                                           80.97                                                                              7.57 × 10.sup.-6            __________________________________________________________________________     *The structure of the substance is characterised by NMR spectroscopy     

                  TABLE 18                                                        ______________________________________                                         ##STR246##                                                                                        Salt         %                                           No.  R.sup.1         form   Mp.   Hem.  IC.sub.50                             ______________________________________                                         ##STR247##      OX     amor- phous*                                                                          41.8                                           ##STR248##      OX     amor- phous*                                                                          24.3                                           ##STR249##      OX     amor- phous*                                                                          73.7                                           ##STR250##      OX     amor- phous*                                                                          53.1                                           ##STR251##      OX     amor- phous*                                           ##STR252##      OX     amor- phous*                                                                          68.22                                          ##STR253##      OX     amor- phous*                                           ##STR254##      BS     amor- phous*                                           ##STR255##      OX     amor- phous*                                           ##STR256##      BS     amor- phous*                                           ##STR257##      OX     amor- phous*                                           ##STR258##      OX     amor- phous*                                                                          30.79                                          ##STR259##      OX     amor- phous*                                           ##STR260##      OX     amor- phous*                                           ##STR261##      OX     amor- phous*                                                                          54.30                                          ##STR262##      OX     amor- phous*                                                                          61.3                                           ##STR263##      Cl     amor- phous*                                                                          5.81                                           ##STR264##      BS     amor- phous*                                           ##STR265##             amor- phous*                                                                          56.0                                           ##STR266##      OX     amor- phous*                                           ##STR267##      OX     amor- phous*                                          ______________________________________                                         *: The structure of the substance is characterised by NMR spectroscopy   

                                      TABLE 19                                    __________________________________________________________________________     ##STR268##                  OX % Hem:                                                                          amorphous* 50.58                            __________________________________________________________________________     *: The structure of the substance is characterised by NMR spectroscopy   

                                      TABLE 20                                    __________________________________________________________________________     ##STR269##                                                                   No.                                                                              R.sup.a R.sup.b                                                                            NR.sup.3 R.sup.4   Salt form                                                                          Mp.  % H                                                                              IC.sub.50                     __________________________________________________________________________    HO         CH.sub.3 O                                                                          ##STR270##        MS   163-169                                                                            86.70                                                                            3.47 × 10.sup.-6        Ph-CH.sub.2 O                                                                            CH.sub.3 O                                                                         "                  MS   amorph*                                                                            31.78                            Ph-CH.sub.2 O                                                                            CH.sub.3 O                                                                         NHCH.sub.2 CH.sub.2 C(CH.sub.3).sub.3                                                            OX   amorph*                                                                            61.85                            Ph-CH.sub.2 O                                                                            Cl                                                                                  ##STR271##        OX   130-132                                                                            32.9                             CH.sub.2 O Cl   "                  OX   143-145                                                                            44.65                            Ph-CH.sub.2 O                                                                            CH.sub.3                                                                           "                  OX   156-157                                                                            49.03                            __________________________________________________________________________     *The structure of the substance is characterised by NMR spectroscopy     

The present invention further relates to the use of the above-mentionedcompounds for the production of agents for treating chronic inflammatoryprocesses, ulcerative colitis and Crohn's disease and for producingagents having an antiproliferative effect. The activity of the compoundscan be explained by their inhibition of the unselective cation channels(UCC).

The pathophysiology of chronic bronchial asthma is based on inflammatoryprocesses which are mediated by the activation of inflammatory cells.(BARNES, 1987; SEIFERT and SCHULTZ, 1991).

The receptor-regulated activation of inflammatory cells (e.g.neutrophilic granulocytes and mast cells or the permanent cell linesHL-60 cells or sensitised RBL cells, i.e. those charged withgammaglobulin E) is inhibited, irrespective of the nature of thestimulating agonists (e.g. endothelin, PAF, leukotrienes, chemotacticalpeptide fMLP or antigen against sensitised mast cells) by blockers ofunselective cation channels (UCC) (RINK, 1990). Through these channelsextracellular calcium, which is responsible for the persistence ofreceptor-mediated cell activations, enters the cells (PUTNEY, 1990). Ifthis supply of calcium is interrupted this results in a blockade of theactivation of inflammatory cells.

Conventional calcium antagonists of the dihydropyridine orphenylalkylamine type do not inhibit either UCCs or inflammatoryprocesses (WELLS et al., 1986).

As a measurement of the cell activation or as a measurement of theinhibition thereof by UCC blockers, the kinetics of the cytoplasmiccalcium ion concentration in fura-2-charged cells is quantifiedfluorometrically using the method described by GRYNKIEWICZ et al.(1985). This procedure has proved a reliable screening method, withinthe scope of the invention, for detecting UCC blockers.

So-called functional THAPSIGARGIN inhibition has proved suitable for thespecific characterisation of blockers of the unselective cationchannels. THAPSIGARGIN is a tumour promoter described by THASTRUP et al.(Proc. Natl. Acad. Sci. (USA), 87, 2466-2470, 1990) which selectivelyand irreversibly inhibits the Ca²⁺ -ATPase of intracellular IP₃-sensitive Ca²⁺ -storeso Consequently the Ca²⁺ -stores are rapidlydepleted. As described by J. PUTNEY (Calcium, 11, 611-624, 1990) thedepletion of these stores constitutes the physiological stimulation foropening up unselective cation channels in the cell membrane. The resultof this is a massive influx of Na⁺ and Ca²⁺ into the cell. Because ofthese properties, Thapsigargin is suitable as an indirect stimulator foragohist- and IP₃ -independent opening up of the unselective cationchannels.

Within the scope of the present invention the Thapsigargin stimulationof unselective cation channels has been carried out successfully onHL-60 cells (human leukaemia cells), on hippocampal and cortical neuronecells and on RBL-cells (rat basophilic lymphoma cells) and in this waythe existence of these channels in particular cell lines wasdemonstrated.

The cytoplasmic Ca²⁺ concentration ([Ca²⁺ ]_(i)) plays an important partin the cell proliferation and in tumour growth (for a summary see L. R.ZACHARSKI, Journal of Medicine 19: 145-177, 1988). In particular, theCa²⁺ -influx into the cell stimulated by receptor activation withconsecutive inositoltriphosphate-(IP₃ -)-mediation would appear to be ofcrucial importance for oncogenic cell proliferation (U. KIKKAWA and Y.NISHIZUKA, Ann. REV. CELL. BIOL. 2: 149-178, 1986). This mechanism alsoplays a part in the formation of metastases and in "Multi-DrugResistance". (For a summary see the abovementioned mentioned publicationby L. R. ZACHARSKI, J. MED. 19: 145-177, 1980).

This hypothesis is supported by the fact that Thapsigargin, as anindirect stimulator of the unselective cation channels (UCC) not onlyleads to a Ca²⁺ -overload in the cell but is also a highly effectivetumour promoter. (V. THASTRUP et al. Proceedings of the NATL. Acad. Sci:(USA) 87: 2466-2470, 1990).

The blockade of the Ca²⁺ -influx by the UCC leads to normalisation ofthe intracellular Ca-ion concentration and hence to inhibition of tumourgrowth etc.

Conventional calcium antagonists do not inhibit these UCC. It has beenfound, surprisingly, that the compounds according to this inventioninhibit the influx of calcium into the cell through the UCC.

As shown by S. H. MURCH et al. (Lancet 339 : 381-385, 15. Feb. 1992)endothelin I plays an important pathophysiological role in inflammatoryintestinal diseases such as ulcerative colitis and Crohn's disease.Using immunohistochemical methods it has been shown that patients withCrohn's disease in the region of the submucosa and patients withulcerative colitis in the region of the lamina propria of the epitheliumof the large intestine show significantly and greatly increasedconcentrations of endothelin I compared with healthy normal people. Itis assumed that the local secretion of endothelin causes massivevasospasms with consecutive disseminated ischaemia with microinfarctswhich are regarded as the actual cause of the above diseases. Thevasospasmogenic effectiveness of endothelin is explained by a Ca²⁺-overload of vascular myocytes. Endothelin primarily triggers an IP₃-mediated intracellular release of Ca²⁺ which is followed by a massivetransmembranal Ca²⁺ -entry through dihydropyridine-insensitive channels.(M. S. Simonson et al. Clin. Invest. Med. 14: 499-507, 1991; T. Masakai,J. Cardiovasc. Pharmacol. 13:Suppl. 5, S1-S4, 1989; D. W. Hay, R. J.Pharmacol. 100: 383-392, 990). These channels are unselective cationchannels which have also been briefly described as existing in cells ofthe large intestine mucosa. (Chr. Siemer and H. Gogelein, Europ. J.Physiol. 420: 319-328, 1992).

The endothelin-stimulated activation of fura-2-charged human leukaemiacells (HL 60 cells) has proved a suitable screening model for detectingfunctional endothelin antagonists. In conformity with G. GRYNKIEWICZ etal. (J. Biol. Chem. 260:3440-3450, 1985) the intracellular Ca²⁺-concentration in the cytoplasm of HL 60 cells (suspensions) can bemonitored by spectrofluorometry and quantified as a measurement of cellactivation by endothelin. The stimulation was effected by adding 0.1 μMendothelin and could be inhibited in a dosage-dependent manner by meansof the substances according to the invention.

The functional endothelin antagonism of the substances according to theinvention is mediated through a blockade of the unselective cationchannels. Consequently, detection of a functional Thapsigarginantagonismon RBL-hml cells is also a suitable screening method for functionalendothelin antagonists.

Carrying out the investigation:

For screening purposes, fura-2-charged adhesive RBL-hm 1 cells arestimulated with 0.1 μM Thapsigargin in a Ca²⁺ -free free incubationmedium. After 4 minutes, extracellular Ca²⁺ is restored to aconcentration of 1.5 mM and, using the fura-2-fluorescence, theexcessive increase in the cytoplasmic Ca²⁺ -concentration caused by amassive transmembranal Ca²⁺ -entry through unselective cation channelsis recorded.

This entry is to be inhibited solely by unselective cation channelblockers in a dosage-dependent manner. Neither conventional calciumantagonists nor specific blockers of agonists which stimulate the IP₃-turnover are able to inhibit the transmembranal Ca²⁺ -entry triggeredindirectly by Thapsigargin. The compounds of the present invention aredistinguished by their inhibktion of UCC.

The fluorometric calcium measurement in the cytoplasm of individualadhering RBL-hml cells is carried out analogously to the methoddescribed by KUDO and OGURA (1986) for neuronal cells. An AXIOVERT 35fluorescence microscope made by ZEISS is used in conjunction with animaging system made by HAMAMATSU, consisting of the ICMS-imageprocessing system, residual light camera with control unit and imageintensifier DVS 3000.

The kinetics of the cytoplasmic Ca²⁺ -concentration is recordedcontinuously as a concentration/time curve after the cell activationstimulated by Thapsigargin (0.1 μM). The curves of two activated cellcultures are compared in the presence and absence of 10 μM testsubstance. The area under these curves (area under the curve=AUC) isintegrated and recorded as a measurement of cell activation. Theinhibitory potency of the UCC-blockers tested is determined using thefollowing equation: ##EQU1## %H=the percentage inhibition of the calciumentry through unselective cation channels which is stimulated andinhibited by 10 μM of test substance.

AUC_(inh) =area under the curve recorded in the presence of thestimulant plus 10 μM inhibitory test substance.

AUC_(control) =area under the curve which is recorded only after theaddition of the stimulant.

Literature relating to the above explanations:

BARNES P. J., I. W. RODGER and N. C. THOMSON Pathogenesis of asthma, in"ASTHMA, basic mechanisms and clinical management"ED by P. J. BARNES;ACADEMIC PRESS, LONDON, 1988

GRYNKIEWICZ G., M. POENIE and R. Y. TSIEN A new generation of Ca²⁺-indicators with greatly improved fluorescence properties J. BIOL. CHEM.260: 3440-3450, 1985

HIDE, M. and M. A. BEAVEN Calcium influx in a rat mast cell (RBL-2H3)life J. BIOL. CHEM. 266 15221-15229, 1991

KUDO, Y. and A. OGURA Glutamate-induced increase in intracellular Ca²⁺-concentration in isolated hippocampal neurones BR. J. PHARMACOL. 89:191-198, 1986

PUTNEY, J. W., jr. Capacitative Calcium entry revised CELL CALCIUM 11:611-624, 1990

RINK, T. J. Receptor-mediated calcium entry FEBS LETT. 268: 381-385,1990

SEIFERT, R. and G. SCHULTZ The superoxide forming NADPH oxidase ofphagocytes: An enzyme system regulated by multiple mechanism REV.PHYSIOL. BIOCHEM. PHARMACOL., Vol. 117, SPRINGER VERL., 1991

WELLS, E., C. G. JACKSON, S. T. HARPER, J. MANN and R. P. EAOYCharacterization of primate bronchoalveolar mast cells II, inhibition ofhistamine, LTC₄ and PGF₂α release from primate bronchoalveolar mastcells and a comparison with rat peritoneal mast cells J. IMMUNOL. 137:3941-3945, 1986.

Results of measurement:

The percentage inhibition of UCC after Thapsigargin stimulation (0.1 μMThapsigargin) in RBL-hm 1 cells is given. The uniform concentration ofthe test substances is 10⁻⁵ mol).

                  TABLE 21                                                        ______________________________________                                        (Data relating to compounds of Tables 1-10)                                   RBL - hm 1 cells - Thapsigargin (0.1 μM) stimulation                       Compound No.         % Inhibition                                             ______________________________________                                         3                   F 41.91                                                   4                   F 66.53                                                   5                   F 80.16                                                   9                   F 67.59                                                   14                  F 80.16                                                   17                  F 52.53                                                   18                  F 57.53                                                   20                  F 57.38                                                   33                  60.55                                                     36                  F 52.19                                                   37                  F 75.77                                                   38a (oxalate)       52.64                                                     39                  48.62                                                     40                  64.99                                                     41 (Structure I)    F 57.22                                                   42                  84.83                                                     43                  F 48.69                                                   46                  68.06                                                     48                  61.34                                                     83                  66.76                                                     89                  53.38                                                     B                   72.35                                                     C                   63.45                                                     E                   F 81.89                                                   F                   48.01                                                     G                   57.52                                                     H                   57.06                                                     K                   76.31                                                     L                   53.14                                                     O                   63.32                                                     Q                   64.28                                                    101                  59.62                                                    102                  F 80.77                                                  104                  54.67                                                    105                  45.09                                                    106                  F 65.69                                                  108                  44.45                                                    109                  74.18                                                    115                  75.53                                                    132                  F 65.14                                                  133                  68.15                                                    ______________________________________                                    

The activity data (%inhibition and IC₅₀) for other compounds arecontained in Tables 13-20.

The functional antiinflammatory effectiveness can be demonstrated bymeans of the following test:

Individual RBL-2H3-cells (a tumour cell line related to the mast cells)adhering to glass slides are used.

The cultivation and attachment of the RBL-2H3-cells are carried out bythe method described by HIDE and BEAVEN (1991). In order to sensitisethe adhesive RBL-2H3-cells the cells are incubated for 2 hours atambient temperature with a 1:2000 diluted commercial gammaglobulinE-solution against a dinitrophenol-bovine serum albumin complex(DNP-BSA-antigen). The cells are then washed. By the addition of 0.1 mlof DNP-BSA-solution (10 μg/ml) there is a massive immunological cellactivation which is mediated by a cytoplasmic Ca²⁺ -overload. Thefluorometric calcium measurement in the cytoplasm of individual adheringRBL-2H3-cells is carried out analogously to the method described by KUDOand OGURA (1986) for neuronal cells, which is also explainedhereinbefore in this specification.

The comparison used in these investigations is (-10 μM) chromoglycatewhich brings about an approximately 50% inhibition of theantigen-induced cell activation.

In this test the above-mentioned compounds demonstrate % H values whichare comparable with the values specified hereinbefore.

Tests on microcultures of various human tumour cell lines using thetetrazolium assay in order to determine the antiproliferative effect ofthe substances according to the invention surprisingly showed that thecompound tested was 5 to 100 times more potent than the comparisonsubstance Verapamil.

The antiproliferative effectiveness of the test substances wasdetermined by means of the MTT test described by MOSMANN (J. IMMUNOL.METH. 65: 55-63, 1983), DENIZOT et al. (J. IMMUNOL. METH. 89: 271-277,1986) and J. ELIASON et al. (INT. J. CANCER 46: 113-117, 1990).(MTT=[3-(4,5-dimethylthiazol-2-yl)2,5-diphenyl-tetrazolium bromide]produced by CHEMICON Inc. El Segundo, Calif., U.S.A.). This indicator ismetabolised only by living cells with intact mitochondria into a blueformazane product. The following human tumour cell lines were used inour test: A 549 (adenocarcinoma of the lung), A 431 (epidermal carcinomaof the vulva), PC 3 (adenocarcinoma of the prostate), SK BR 3(adenocarcinoma of the breast), HT 29 (CX1 1) (adenocarcinoma of thecolon) and K 562 (chronic myeloid leukaemia cell). The test was carriedout on microtitre plates. Each well contained 100 μl of a cellsuspension (0.2×10⁶ cells per ml). The incubation medium used was RPMI1640 with 10% heat-inactivated foetal calves' serum and 50 μg/ml ofgentamycin. The cell suspensions were incubated for 0, 24, 48 or 72hours in air with a humidity at saturation point in a CO₂ (5%)/air (95%)mixture at 37° C., incubated in the presence and absence of variableconcentrations of antiproliferative test substances. The test substanceswere dissolved in DMSO (final dilution: 0.1%). Then 10 μl ofMTT-solution (3 mg/ml) were added, followed after 3 hours by 100 μl ofan isopropanol solution containing 0.08 N HCl. After a further hour, thelight absorption at 570 nm (comparative wavelength 630 nm) wasdetermined in a microplate reader. The light absorption is directlyproportional to the number of living cells. The half-maximum inhibitoryconcentrations of the substances tested were 1 μg/ml.

The vasospasmolytic effectiveness of the above-mentioned functionalendothelin and Thapsigargin antagonists were confirmed on an isolatedblood vessel preparation: coronary perfusion was continuouslyquantified, on retrogressively perfused, spontaneously beatingLANGENDORFF hearts taken from rats, by means of electromagnetic flowmeasurement (apparatus supplied by Hugo Sachs Elektronik, MARCH). Thismeasuring apparatus could be used to record the extent, duration andpattern of vascular spasms with a high degree of accuracy. If perfusionis carried out with 100 nM endothelin concentration, the coronaryperfusion flow is reduced from 11 to 5 ml/min. The restriction inperfusion can be reversed by means of the substances according to theinvention. The potencies of the compounds according to the inventionwith regard to Thapsigargin inhibition on fura-2-charged RBL-hml-cellsor the effectiveness of endothelin-inhibition on fura-2-charged HL 60cells correlates clearly with the vasospasmolytic effectiveness of thetest substances detected on the Langendorff preparation. It can beconcluded from this that, underlying the vasospasmolytic endothelinantagonism of the substances tested, there is a blockade of theunselective cation channels.

Examples of Pharmaceutical Preparations

    ______________________________________                                        a) Coated tablets                                                             1 tablet core contains:                                                       ______________________________________                                        Active substance of general formula I                                                                  30.0   mg                                            Lactose                  100.0  mg                                            Corn starch              75.0   mg                                            Gelatine                 3.0    mg                                            Magnesium stearate       2.0    mg                                                                     210.0  mg                                            ______________________________________                                    

Preparation

The active substance mixed with lactose and corn starch is granulatedwith a 10% aqueous gelatine solution through a 1 mm mesh screen, driedat 40° C. and rubbed through a screen once more. The granules thusobtained are mixed with magnesium stearate and compressed. The coresproduced in this way are coated in the usual manner with a coatingconsisting of an aqueous suspension of sugar, titanium dioxide, talc andgum arabic. The finished coated tablets are polished with beeswax.

    ______________________________________                                        b) Tablets                                                                    ______________________________________                                        Active substance of general formula I                                                                  30.0   mg                                            Lactose                  100.0  mg                                            Corn starch              70.0   mg                                            Soluble starch           7.0    mg                                            Magnesium stearate       3.0    mg                                                                     210.0  mg                                            ______________________________________                                    

Preparation

The active substance and magnesium stearate are granulated with anaqueous solution of the soluble starch, the granules are dried andintimately mixed with lactose and corn starch. The mixture is thencompressed into tablets weighing 210 mg.

    ______________________________________                                        c) Capsules                                                                   ______________________________________                                        Active substance according to claim 1                                                                  20.0   mg                                            Lactose                  230.0  mg                                            Corn starch              40.0   mg                                            Talc                     10.0   mg                                                                     300.0  mg                                            ______________________________________                                    

Preparation

The active substance, lactose and corn starch are first combined in amixer and then in a grinding machine. The mixture is returned to themixer, thoroughly combined with the talc and mechanically packed intohard gelatine capsules.

    ______________________________________                                        d) Tablets                                                                    ______________________________________                                        Active substance according to the invention                                                             40.0   mg                                           Lactose                   100.0  mg                                           Corn starch               50.0   mg                                           Colloidal silica          2.0    mg                                           Magnesium stearate        3.0    mg                                           Total                     200.0  mg                                           ______________________________________                                    

Preparation

The active substance is mixed with some of the excipients and granulatedwith a solution of the soluble starch in water. After the granules havebeen dried the remaining excipients are added and the mixture iscompressed to form tablets.

    ______________________________________                                        e) Coated tablets                                                             ______________________________________                                        Active substance according to the invention                                                             20.0   mg                                           Lactose                   100.0  mg                                           Corn starch               65.0   mg                                           Colloidal silica          2.0    mg                                           Soluble starch            5.0    mg                                           Magnesium stearate        3.0    mg                                           Total                     195.0  mg                                           ______________________________________                                    

Preparation

The active substance and excipients are compressed to form tablet coresas described in Example 1 which are then coated with sugar, talc and gumarabic in the usual way.

    ______________________________________                                        f) Suppositories                                                              ______________________________________                                        Active substance according to the invention                                                             50.0   mg                                           Lactose                   250.0  mg                                           Suppository mass q.s. ad  1.7    g                                            ______________________________________                                    

Preparation

The active substance and lactose are mixed together and the mixture isuniformly suspended in the molten suppository mass. The suspensions arepoured into cooled moulds to form suppositories weighing 1.7 g.

    ______________________________________                                        g) Ampoules                                                                   ______________________________________                                        Active substance according to the invention                                                             20.0   mg                                           Sodium chloride           5.0    mg                                           Twice distilled water q.s. ad                                                                           2.0    ml                                           ______________________________________                                    

Preparation

The active substance and sodium chloride are dissolved in twicedistilled water and the solution is transferred under sterile conditionsinto ampoules.

    ______________________________________                                        h) Ampoules                                                                   ______________________________________                                        Active substance according to the invention                                                             10.0   mg                                           Sodium chloride           7.0    mg                                           Twice distilled water q.s. ad                                                                           1.0    mg                                           ______________________________________                                    

    ______________________________________                                        i) Drops                                                                      ______________________________________                                        Active substance according to the invention                                                             0.70    g                                           Methyl p-hydroxybenzoate  0.07    g                                           Propyl p-hydroxybenzoate  0.03    g                                           Demineralised water q.s. ad                                                                             100.00  ml                                          ______________________________________                                    

Preparation

The active substance and preservatives are dissolved in demineralisedwater, the solution is filtered and transferred into vials holding 100ml.

What is claimed is:
 1. A compound of formula ##STR272## wherein A is abenzo;mis 2; R² are in positions 6 and 7 and independently of each othermay be hydroxy, (C₁₋₄)alkoxy, benzyloxy, halogen, (C₁₋₄)alkyl,methanesulphonyloxy or methanesulphonamido or two adjacent substituentsR² together may be --O--CH₂ --O or --O--CH₂ --CH₂ --O--; R¹ is(C₄₋₆)cyloalkyl or (C₄₋₆)cycloalkyl(C₁₋₅)-alkyl; R³ and R⁴ independentlyor each other are(a) hydrogen, (b) branched or unbranched C₃₋₆ -alkenyl,(c) branched or unbranched C₃₋₆ -alkynyl, (d) branched or unbranchedC₁₋₁₂ -alkyl, whilst the alkyl may be substituted byhydroxy,(C₁₋₄)alkoxy, di(C₁₋₄)akylamino, furyl, pyridyl, pyrrolidinyl,N-methylpyrrolidinyl, morpholino, indolyl, nitrilo, thienyl, adamantyl,cyclohexyl, phenoxy, napthyloxy or phenyl, whilst this phenyl or thephenyl contained in the phenoxy group may be mono-, di- ortrisubstituted by hydroxy, (C₁₋₄)alkoxy, benzyloxy, halogen, CF₃, N₃,(C₁₋₄)alkyl, adamantyl, --SO₂ NH₂ or by the bridge --O--CH₂ --O--; or R³is hydrogen and R⁴ is cyclohexyl, phenyl, fluorophenyl, pyridyl orN-benzylpiperidyl; or R³ and R⁴ together with the nitrogen atom to whichthey arebound represent pyrrolidinyl, piperidinyl, morpholino,thiomorpholino, the group ##STR273## or piperazinyl, whilst thepiperazinyl ring may optionally be N-substituted by methyl,unsubstituted phenyl, mono- or di(C₁₋₄)alkoxyphenyl, pyrimidinyl,phenyl(C₁₋₄)alkyl or ##STR274## or the salts thereof withphysiologically acceptable acids, bases or complexing agents.
 2. Thecompounds as recited in claim 1, wherein R¹ is cyclohexyl,cyclohexylmethylene or cyclobutylmethylene.
 3. The compounds as recitedin claim 2 wherein NR³ R⁴ is ##STR275##
 4. The compounds as recited inclaim 1 of formula ##STR276##
 5. The compounds as recited in claim 1wherein NR³ R⁴ is ##STR277##
 6. A method for treating chronicinflammatory processes, ulcerative colitis and Crohn's disease in awarm-blooded animal, which comprises administering to said animal atherapeutically effective amount of the compound of claim
 1. 7. A methodfor treating diseases in a warm-blooded animal that responds to an agentwith antiproliferative activity which comprises administering to saidanimal a therapeutically effective amount of the compound of claim 1.